Reece R J, Rickles R J, Ptashne M
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Gene. 1993 Apr 15;126(1):105-7. doi: 10.1016/0378-1119(93)90596-u.
A DNA fragment encoding the yeast GAL4 transcriptional activator DNA-binding domain (amino acids 1-93) was cloned into an Escherichia coli expression vector such that the overproduced protein is tagged with six histidine residues and a factor Xa protease cleavage site. The vector also contains unique restriction sites at the 3' end of the gene to allow the construction of fusion proteins. These fusion proteins can easily be purified to homogeneity and their activity tested in vitro.
将编码酵母GAL4转录激活因子DNA结合结构域（氨基酸1 - 93）的DNA片段克隆到大肠杆菌表达载体中，使得过量表达的蛋白质带有六个组氨酸残基和一个因子Xa蛋白酶切割位点。该载体在基因的3'端还含有独特的限制性酶切位点，以允许构建融合蛋白。这些融合蛋白可以很容易地纯化至同质，并在体外测试其活性。
